RESUMEN
Most sensory information destined for the neocortex is relayed through the thalamus, where considerable transformation occurs1,2. One means of transformation involves interactions between excitatory thalamocortical neurons that carry data to the cortex and inhibitory neurons of the thalamic reticular nucleus (TRN) that regulate the flow of those data3-6. Although the importance of the TRN has long been recognised7-9, understanding of its cell types, their organization and their functional properties has lagged behind that of the thalamocortical systems they control. Here we address this by investigating the somatosensory and visual circuits of the TRN in mice. In the somatosensory TRN we observed two groups of genetically defined neurons that are topographically segregated and physiologically distinct, and that connect reciprocally with independent thalamocortical nuclei through dynamically divergent synapses. Calbindin-expressing cells-located in the central core-connect with the ventral posterior nucleus, the primary somatosensory thalamocortical relay. By contrast, somatostatin-expressing cells-which reside along the surrounding edges of the TRN-synapse with the posterior medial thalamic nucleus, a higher-order structure that carries both top-down and bottom-up information10-12. The two TRN cell groups process their inputs in pathway-specific ways. Synapses from the ventral posterior nucleus to central TRN cells transmit rapid excitatory currents that depress deeply during repetitive activity, driving phasic spike output. Synapses from the posterior medial thalamic nucleus to edge TRN cells evoke slower, less depressing excitatory currents that drive more persistent spiking. Differences in the intrinsic physiology of TRN cell types, including state-dependent bursting, contribute to these output dynamics. The processing specializations of these two somatosensory TRN subcircuits therefore appear to be tuned to the signals they carry-a primary central subcircuit tuned to discrete sensory events, and a higher-order edge subcircuit tuned to temporally distributed signals integrated from multiple sources. The structure and function of visual TRN subcircuits closely resemble those of the somatosensory TRN. These results provide insights into how subnetworks of TRN neurons may differentially process distinct classes of thalamic information.
Asunto(s)
Vías Nerviosas , Núcleos Talámicos/citología , Núcleos Talámicos/fisiología , Potenciales de Acción , Animales , Calbindinas/metabolismo , Potenciales Evocados Somatosensoriales , Potenciales Evocados Visuales , Femenino , Cinética , Masculino , Ratones , Inhibición Neural , Neuronas/metabolismo , Somatostatina/metabolismo , Sinapsis/metabolismoRESUMEN
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
RESUMEN
Knowledge of thalamocortical (TC) processing comes mainly from studying core thalamic systems that project to middle layers of primary sensory cortices. However, most thalamic relay neurons comprise a matrix of cells that are densest in the "nonspecific" thalamic nuclei and usually target layer 1 (L1) of multiple cortical areas. A longstanding hypothesis is that matrix TC systems are crucial for regulating neocortical excitability during changing behavioral states, yet we know almost nothing about the mechanisms of such regulation. It is also unclear whether synaptic and circuit mechanisms that are well established for core sensory TC systems apply to matrix TC systems. Here we describe studies of thalamic matrix influences on mouse prefrontal cortex using optogenetic and in vitro electrophysiology techniques. Channelrhodopsin-2 was expressed in midline and paralaminar (matrix) thalamic neurons, and their L1-projecting TC axons were activated optically. Contrary to conventional views, we found that matrix TC projections to L1 could transmit relatively strong, fast, high-fidelity synaptic signals. L1 TC projections preferentially drove inhibitory interneurons of L1, especially those of the late-spiking subtype, and often triggered feedforward inhibition in both L1 interneurons and pyramidal cells of L2/L3. Responses during repetitive stimulation were far more sustained for matrix than for core sensory TC pathways. Thus, matrix TC circuits appear to be specialized for robust transmission over relatively extended periods, consistent with the sort of persistent activation observed during working memory and potentially applicable to state-dependent regulation of excitability.
Asunto(s)
Corteza Prefrontal/fisiología , Tálamo/fisiología , Animales , Estimulación Eléctrica/métodos , Potenciales Postsinápticos Excitadores/fisiología , Interneuronas/fisiología , Ratones , Ratones Endogámicos ICR , Imagen Molecular/métodos , Inhibición Neural/fisiología , Vías Nerviosas/fisiología , Optogenética/métodos , Corteza Prefrontal/anatomía & histología , Células Piramidales/fisiologíaRESUMEN
The thalamus provides fundamental input to the neocortex. This input activates inhibitory interneurons more strongly than excitatory neurons, triggering powerful feedforward inhibition. We studied the mechanisms of this selective neuronal activation using a mouse somatosensory thalamocortical preparation. Notably, the greater responsiveness of inhibitory interneurons was not caused by their distinctive intrinsic properties but was instead produced by synaptic mechanisms. Axons from the thalamus made stronger and more frequent excitatory connections onto inhibitory interneurons than onto excitatory cells. Furthermore, circuit dynamics allowed feedforward inhibition to suppress responses in excitatory cells more effectively than in interneurons. Thalamocortical excitatory currents rose quickly in interneurons, allowing them to fire action potentials before significant feedforward inhibition emerged. In contrast, thalamocortical excitatory currents rose slowly in excitatory cells, overlapping with feedforward inhibitory currents that suppress action potentials. These results demonstrate the importance of selective synaptic targeting and precise timing in the initial stages of neocortical processing.
Asunto(s)
Neocórtex/citología , Inhibición Neural/fisiología , Vías Nerviosas/fisiología , Neuronas/fisiología , Sinapsis/fisiología , Tálamo/fisiología , Animales , Animales Recién Nacidos , Relación Dosis-Respuesta en la Radiación , Estimulación Eléctrica/métodos , Potenciales Postsinápticos Excitadores/fisiología , Potenciales Postsinápticos Excitadores/efectos de la radiación , Proteínas Fluorescentes Verdes/biosíntesis , Técnicas In Vitro , Potenciales de la Membrana/fisiología , Potenciales de la Membrana/efectos de la radiación , Ratones , Ratones Endogámicos ICR , Ratones Transgénicos , Técnicas de Placa-ClampRESUMEN
Gap junctions mediate metabolic and electrical interactions between some cells of the CNS. For many types of neurons, gap junction-mediated electrical coupling is most prevalent during early development, then decreases sharply with maturation. However, neurons in the thalamic reticular nucleus (TRN), which exert powerful inhibitory control over thalamic relay cells, are electrically coupled in relatively mature animals. It is not known whether TRN cells or any neurons that are electrically coupled when mature are also coupled during early development. We used dual whole-cell recordings in mouse brain slices to study the postnatal development of electrical and chemical synapses that interconnect TRN neurons. Inhibitory chemical synapses were seen as early as postnatal day 4 but were infrequent at all ages, whereas TRN cells were extensively connected by electrical synapses from birth onward. Surprisingly, the functional strength of electrical coupling, assayed under steady-state conditions or during spiking, remained relatively constant as the brain matured despite dramatic concurrent changes of intrinsic membrane properties. Most notably, neuronal input resistances declined almost eightfold during the first two postnatal weeks, but there were offsetting increases in gap junctional conductances. This suggests that the size or number of gap junctions increase homeostatically to compensate for leakier nonjunctional membranes. Additionally, we found that the ability of electrical synapses to synchronize high frequency subthreshold signals improved as TRN cells matured. Our results demonstrate that certain central neurons may maintain or even increase their gap junctional communication as they mature.
Asunto(s)
Sinapsis Eléctricas/fisiología , Uniones Comunicantes/fisiología , Neuronas/fisiología , Tálamo/crecimiento & desarrollo , Tálamo/fisiología , Potenciales de Acción , Envejecimiento , Animales , Animales Recién Nacidos , Membrana Celular/fisiología , Estimulación Eléctrica , Técnicas In Vitro , Potenciales de la Membrana , Ratones , Técnicas de Placa-ClampRESUMEN
Gap junction-mediated electrical synapses interconnect diverse types of neurons in the mammalian brain, and they may play important roles in the synchronization and development of neural circuits. Thalamic relay neurons are the major source of input to neocortex. Electrical synapses have not been directly observed between relay neurons in either developing or adult animals. We tested for electrical synapses by recording from pairs of relay neurons in acute slices of developing ventrobasal nucleus (VBN) of the thalamus from rats and mice. Electrical synapses were common between VBN relay neurons during the first postnatal week, and then declined sharply during the second week. Electrical coupling was reduced among cells of connexin36 (Cx36) knockout mice; however, some neuron pairs remained coupled. This implies that electrical synapses between the majority of coupled VBN neurons require Cx36 but that other gap junction proteins also contribute. The anatomical distribution of a beta-galactosidase reporter indicated that Cx36 was expressed in some VBN neurons during the first postnatal week and sharply declined over the second week, consistent with our physiological results. VBN relay neurons also communicated via chemical synapses. Rare pairs of relay neurons excited one another monosynaptically. Much more commonly, spikes in one relay neuron evoked disynaptic inhibition (via the thalamic reticular nucleus) in the same or a neighbouring relay neuron. Disynaptic inhibition between VBN cells emerged as electrical coupling was decreasing, during the second postnatal week. Our results demonstrate that thalamic relay neurons communicate primarily via electrical synapses during early postnatal development, and then lose their electrical coupling as a chemical synapse-mediated inhibitory circuit matures.
Asunto(s)
Neuronas/fisiología , Sinapsis/fisiología , Tálamo/fisiología , Envejecimiento/fisiología , Animales , Conexinas/genética , Electrofisiología , Inmunohistoquímica , Potenciales de la Membrana/fisiología , Ratones , Ratones Noqueados , Red Nerviosa/química , Red Nerviosa/fisiología , Conducción Nerviosa/fisiología , Neurotransmisores/fisiología , Ratas , Ratas Sprague-Dawley , Tálamo/citología , Tálamo/crecimiento & desarrollo , beta-Galactosidasa/metabolismo , Proteína delta-6 de Union ComunicanteRESUMEN
The rodent somatosensory cortex includes well-defined examples of cortical columns-the barrel columns-that extend throughout the cortical depth and are defined by discrete clusters of neurons in layer 4 (L4) called barrels. Using the cell-type-specific Ntsr1-Cre mouse line, we found that L6 contains infrabarrels, readily identifiable units that align with the L4 barrels. Corticothalamic (CT) neurons and their local axons cluster within the infrabarrels, whereas corticocortical (CC) neurons are densest between infrabarrels. Optogenetic experiments showed that CC cells received robust input from somatosensory thalamic nuclei, whereas CT cells received much weaker thalamic inputs. We also found that CT neurons are intrinsically less excitable, revealing that both synaptic and intrinsic mechanisms contribute to the low firing rates of CT neurons often reported in vivo. In summary, infrabarrels are discrete cortical circuit modules containing two partially separated excitatory networks that link long-distance thalamic inputs with specific outputs.
Asunto(s)
Vías Nerviosas/fisiología , Neuronas/fisiología , Corteza Somatosensorial/fisiología , Tálamo/fisiología , Vibrisas/fisiología , Animales , Recuento de Células , Ratones , Ratones Transgénicos , Vías Nerviosas/ultraestructura , Neuronas/clasificación , Neuronas/ultraestructura , Corteza Somatosensorial/ultraestructura , Tálamo/ultraestructura , Vibrisas/citologíaRESUMEN
Synchronous activity is common in the neocortex, although its significance, mechanisms, and development are poorly understood. Previous work showed that networks of electrically coupled inhibitory interneurons called low-threshold spiking (LTS) cells can fire synchronously when stimulated by metabotropic glutamate receptors. Here we found that the coordinated inhibition emerging from an activated LTS network could induce correlated spiking patterns among neighboring excitatory cells. Synchronous activity among LTS cells was absent at postnatal day 12 (P12) but appeared abruptly over the next few days. The rapid development of the LTS-synchronizing system coincided with the maturation of the inhibitory outputs and intrinsic membrane properties of the neurons. In contrast, the incidence and magnitude of electrical synapses remained constant between P8 and P15. The developmental transformation of LTS interneurons into a synchronous, oscillatory network overlaps with the onset of active somatosensory exploration, suggesting a potential role for this synchronizing system in sensory processing.
Asunto(s)
Potenciales de Acción/fisiología , Envejecimiento/fisiología , Animales Recién Nacidos/crecimiento & desarrollo , Sincronización Cortical , Neocórtex/fisiología , Animales , Técnicas In Vitro , Interneuronas/fisiología , Neocórtex/crecimiento & desarrollo , Red Nerviosa/crecimiento & desarrollo , Inhibición Neural/fisiología , Ratas , Ratas Sprague-Dawley , Tiempo de Reacción/fisiología , Sinapsis/fisiologíaAsunto(s)
Interneuronas/fisiología , Neocórtex/fisiología , Potenciales Postsinápticos Excitadores/fisiología , Humanos , Interneuronas/ultraestructura , Microscopía Electrónica , Neocórtex/citología , Neocórtex/ultraestructura , Red Nerviosa/fisiología , Neurópilo/fisiología , Neurópilo/ultraestructura , Terminales Presinápticos/fisiología , Terminales Presinápticos/ultraestructura , Células Piramidales/fisiología , Células Piramidales/ultraestructura , Sinapsis/fisiología , Sinapsis/ultraestructuraRESUMEN
Corticothalamic neurons provide massive input to the thalamus. This top-down projection may allow the cortex to regulate sensory processing by modulating the excitability of thalamic cells. Layer 6 corticothalamic neurons monosynaptically excite thalamocortical cells, but also indirectly inhibit them by driving inhibitory cells of the thalamic reticular nucleus. Whether corticothalamic activity generally suppresses or excites the thalamus remains unclear. Here we show that the corticothalamic influence is dynamic, with the excitatory-inhibitory balance shifting in an activity-dependent fashion. During low-frequency activity, corticothalamic effects are mainly suppressive, whereas higher-frequency activity (even a short bout of gamma frequency oscillations) converts the corticothalamic influence to enhancement. The mechanism of this switching depends on distinct forms of short-term synaptic plasticity across multiple corticothalamic circuit components. Our results reveal an activity-dependent mechanism by which corticothalamic neurons can bidirectionally switch the excitability and sensory throughput of the thalamus, possibly to meet changing behavioral demands.
Asunto(s)
Potenciales Postsinápticos Excitadores/fisiología , Vías Nerviosas/fisiología , Corteza Somatosensorial/fisiología , Sinapsis/fisiología , Tálamo/fisiología , Animales , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Estimulantes del Sistema Nervioso Central/farmacología , Channelrhodopsins , Antagonistas de Aminoácidos Excitadores/farmacología , Técnicas In Vitro , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Ratones , Ratones Endogámicos ICR , Ratones Transgénicos , N-Metilaspartato/farmacología , Optogenética , Picrotoxina/farmacología , Receptores de Neurotensina/genética , Receptores de Neurotensina/metabolismo , Sinapsis/genética , Valina/análogos & derivados , Valina/farmacología , Ácido gamma-Aminobutírico/farmacologíaRESUMEN
Thalamocortical and corticothalamic pathways mediate bidirectional communication between the thalamus and neocortex. These pathways are entwined, making their study challenging. Here we used lentiviruses to express channelrhodopsin-2 (ChR2), a light-sensitive cation channel, in either thalamocortical or corticothalamic projection cells. Infection occurred only locally, but efferent axons and their terminals expressed ChR2 strongly, allowing selective optical activation of each pathway. Laser stimulation of ChR2-expressing thalamocortical axons/terminals evoked robust synaptic responses in cortical excitatory cells and fast-spiking (FS) inhibitory interneurons, but only weak responses in somatostatin-containing interneurons. Strong FS cell activation led to feedforward inhibition in all cortical neuron types, including FS cells. Corticothalamic stimulation excited thalamic relay cells and inhibitory neurons of the thalamic reticular nucleus (TRN). TRN activation triggered inhibition in relay cells but not in TRN neurons. Thus, a major difference between thalamocortical and corticothalamic processing was the extent to which feedforward inhibitory neurons were themselves engaged by feedforward inhibition.
Asunto(s)
Axones/fisiología , Neocórtex/fisiología , Red Nerviosa/fisiología , Fenómenos Ópticos , Transducción de Señal/fisiología , Tálamo/fisiología , Animales , Rayos Láser , Ratones , Inhibición Neural/fisiologíaRESUMEN
Electrical synapses are composed of gap junction channels that interconnect neurons. They occur throughout the mammalian brain, although this has been appreciated only recently. Gap junction channels, which are made of proteins called connexins, allow ionic current and small organic molecules to pass directly between cells, usually with symmetrical ease. Here we review evidence that electrical synapses are a major feature of the inhibitory circuitry in the thalamocortical system. In the neocortex, pairs of neighboring inhibitory interneurons are often electrically coupled, and these electrical connections are remarkably specific. To date, there is evidence that five distinct subtypes of inhibitory interneurons in the cortex make electrical interconnections selectively with interneurons of the same subtype. Excitatory neurons (i.e., pyramidal and spiny stellate cells) of the mature cortex do not appear to make electrical synapses. Within the thalamus, electrical coupling is observed in the reticular nucleus, which is composed entirely of GABAergic neurons. Some pairs of inhibitory neurons in the cortex and reticular thalamus have mixed synaptic connections: chemical (GABAergic) inhibitory synapses operating in parallel with electrical synapses. Inhibitory neurons of the thalamus and cortex express the gap junction protein connexin 36 (C x 36), and knocking out its gene abolishes nearly all of their electrical synapses. The electrical synapses of the thalamocortical system are strong enough to mediate robust interactions between inhibitory neurons. When pairs or groups of electrically coupled cells are excited by synaptic input, receptor agonists, or injected current, they typically display strong synchrony of both subthreshold voltage fluctuations and spikes. For example, activating metabotropic glutamate receptors on coupled pairs of cortical interneurons or on thalamic reticular neurons can induce rhythmic action potentials that are synchronized with millisecond precision. Electrical synapses offer a uniquely fast, bidirectional mechanism for coordinating local neural activity. Their widespread distribution in the thalamocortical system suggests that they serve myriad functions. We are far from a complete understanding of those functions, but recent experiments suggest that electrical synapses help to coordinate the temporal and spatial features of various forms of neural activity.
Asunto(s)
Corteza Cerebral/fisiología , Conexinas/fisiología , Uniones Comunicantes/fisiología , Interneuronas/fisiología , Tálamo/fisiología , Animales , Comunicación Celular/fisiología , Corteza Cerebral/ultraestructura , Uniones Comunicantes/ultraestructura , Humanos , Interneuronas/ultraestructura , Inhibición Neural/fisiología , Vías Nerviosas/fisiología , Vías Nerviosas/ultraestructura , Transmisión Sináptica/fisiología , Tálamo/ultraestructuraRESUMEN
To facilitate an understanding of auditory thalamocortical mechanisms, we have developed a mouse brain-slice preparation with a functional connection between the ventral division of the medial geniculate (MGv) and the primary auditory cortex (ACx). Here we present the basic characteristics of the slice in terms of physiology (intracellular and extracellular recordings, including current source density analysis), pharmacology (including glutamate receptor involvement), and anatomy (gross anatomy, Nissl, parvalbumin immunocytochemistry, and tract tracing with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate). Thalamocortical transmission in this preparation (the "primary" slice) involves both alpha-amino-3-hydroxy-5-methylisoxazole-4-proprionic acid/kainate and N-methyl-D-aspartate-type glutamate receptors that appear to mediate monosynaptic inputs to layers 3-4 of ACx. MGv stimulation also initiates disynaptic inhibitory postsynaptic potentials and longer-duration intracortical, polysynaptic activity. Important differences between responses elicited by MGv versus conventional columnar ("on-beam") stimulation emphasize the necessity of thalamic activation to infer thalamocortical mechanisms. We also introduce a second slice preparation, the "shell" slice, obtained from the brain region immediately ventral to the primary slice, that may contain a nonprimary thalamocortical pathway to temporal cortex. In the shell slice, stimulation of the thalamus or the region immediately ventral to it appears to produce fast activation of synapses in cortical layer 1 followed by robust intracortical polysynaptic activity. The layer 1 responses may result from orthodromic activation of nonprimary thalamocortical pathways; however, a plausible alternative could involve antidromic activation of corticotectal neurons and their layer 1 collaterals. The primary and shell slices will provide useful tools to investigate mechanisms of information processing in the ACx.
Asunto(s)
Corteza Auditiva/fisiología , Vías Auditivas/fisiología , Transmisión Sináptica/fisiología , Tálamo/fisiología , Potenciales de Acción/fisiología , Animales , Corteza Auditiva/anatomía & histología , Corteza Auditiva/efectos de los fármacos , Vías Auditivas/anatomía & histología , Vías Auditivas/efectos de los fármacos , Calcio/metabolismo , Calcio/farmacología , Estimulación Eléctrica , Antagonistas de Aminoácidos Excitadores/farmacología , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Potenciales Postsinápticos Excitadores/fisiología , Colorantes Fluorescentes , Cuerpos Geniculados/anatomía & histología , Cuerpos Geniculados/efectos de los fármacos , Cuerpos Geniculados/fisiología , Técnicas In Vitro , Ratones , Ratones Endogámicos , Inhibición Neural/fisiología , Tiempo de Reacción/fisiología , Receptores AMPA/antagonistas & inhibidores , Receptores de N-Metil-D-Aspartato/antagonistas & inhibidores , Transmisión Sináptica/efectos de los fármacos , Tálamo/anatomía & histología , Tálamo/efectos de los fármacosRESUMEN
Recently, great interest has been shown in understanding the functional roles of specific gap junction proteins (connexins) in brain, lens, retina, and elsewhere. Some progress has been made by studying knockout mice with targeted connexin deletions. For example, such studies have implicated the gap junction protein Cx36 in synchronizing rhythmic activity of neurons in several brain regions. Although knockout strategies are informative, they can be problematic, because compensatory changes sometimes occur during development. Therefore, it would be extremely useful to have pharmacological agents that block specific connexins, without major effects on other gap junctions or membrane channels. We show that mefloquine, an antimalarial drug, is one such agent. It blocked Cx36 channels, expressed in transfected N2A neuroblastoma cells, at low concentrations (IC(50) approximately 300 nM). Mefloquine also blocked channels formed by the lens gap junction protein, Cx50 (IC(50) approximately 1.1 microM). However, other gap junctions (e.g., Cx43, Cx32, and Cx26) were only affected at concentrations 10- to 100-fold higher. To further examine the utility and specificity of this compound, we characterized its effects in acute brain slices. Mefloquine, at 25 microM, blocked gap junctional coupling between interneurons in neocortical slices, with minimal nonspecific actions. At this concentration, the only major side effect was an increase in spontaneous synaptic activity. Mefloquine (25 microM) caused no significant change in evoked excitatory or inhibitory postsynaptic potentials, and intrinsic cellular properties were also mostly unaffected. Thus, mefloquine is expected to be a useful tool to study the functional roles of Cx36 and Cx50.